国际泌尿系统杂志
國際泌尿繫統雜誌
국제비뇨계통잡지
INTERNATIONAL JOURNAL OF UROLOGY AND NEPHROLOGY
2008年
6期
732-736
,共5页
干细胞%骨髓细胞
榦細胞%骨髓細胞
간세포%골수세포
Stem Cells%Bone Marrow Cells
目的 观察贴壁法分离提取大鼠乳鼠骨髓间充质干细胞(mesenchymal stem cells,MSCs)并进行体外扩增培养的可行性并探讨其生物学特性,为进一步实验研究作基础.方法 实验2008-01/06于南昌大学第一附属医院泌尿外科研究所完成.贴壁法分离培养SD大鼠乳鼠MSCs,经CD29、CD34、CD44、CD45鉴定MSCs,并绘制不同代数细胞生长曲线和贴壁率曲线分析其生物学特性.结果 ①原代细胞生长潜伏期略长,孵育24 h后小部分圆形单核细胞开始贴壁,48 h可见巢状克隆集落形成,贴壁细胞呈多形性,贴壁3d后增殖加快,9d后基本融合.②生长曲线及贴壁率曲线提示第2至第6代细胞接种存活率基本相同,生长曲线基本一致,而第7代以后细胞的生长速度渐缓,贴壁时间延长,细胞接种存活率也明显降低,并出现衰化现象.③免疫组化鉴定结果 显示CD29、CD44表达基本阳性,而CD34、CD45表达阴性.结论 贴壁法比较容易分离培养MSCs,为组织工程的应用提供了足够的种子来源,MSCs有一定的增殖能力但并不是无限的,而是随传代扩增逐渐衰化,因此了解和掌握MSCs的生物学特性为进一步研究应用打下基础.
目的 觀察貼壁法分離提取大鼠乳鼠骨髓間充質榦細胞(mesenchymal stem cells,MSCs)併進行體外擴增培養的可行性併探討其生物學特性,為進一步實驗研究作基礎.方法 實驗2008-01/06于南昌大學第一附屬醫院泌尿外科研究所完成.貼壁法分離培養SD大鼠乳鼠MSCs,經CD29、CD34、CD44、CD45鑒定MSCs,併繪製不同代數細胞生長麯線和貼壁率麯線分析其生物學特性.結果 ①原代細胞生長潛伏期略長,孵育24 h後小部分圓形單覈細胞開始貼壁,48 h可見巢狀剋隆集落形成,貼壁細胞呈多形性,貼壁3d後增殖加快,9d後基本融閤.②生長麯線及貼壁率麯線提示第2至第6代細胞接種存活率基本相同,生長麯線基本一緻,而第7代以後細胞的生長速度漸緩,貼壁時間延長,細胞接種存活率也明顯降低,併齣現衰化現象.③免疫組化鑒定結果 顯示CD29、CD44錶達基本暘性,而CD34、CD45錶達陰性.結論 貼壁法比較容易分離培養MSCs,為組織工程的應用提供瞭足夠的種子來源,MSCs有一定的增殖能力但併不是無限的,而是隨傳代擴增逐漸衰化,因此瞭解和掌握MSCs的生物學特性為進一步研究應用打下基礎.
목적 관찰첩벽법분리제취대서유서골수간충질간세포(mesenchymal stem cells,MSCs)병진행체외확증배양적가행성병탐토기생물학특성,위진일보실험연구작기출.방법 실험2008-01/06우남창대학제일부속의원비뇨외과연구소완성.첩벽법분리배양SD대서유서MSCs,경CD29、CD34、CD44、CD45감정MSCs,병회제불동대수세포생장곡선화첩벽솔곡선분석기생물학특성.결과 ①원대세포생장잠복기략장,부육24 h후소부분원형단핵세포개시첩벽,48 h가견소상극륭집락형성,첩벽세포정다형성,첩벽3d후증식가쾌,9d후기본융합.②생장곡선급첩벽솔곡선제시제2지제6대세포접충존활솔기본상동,생장곡선기본일치,이제7대이후세포적생장속도점완,첩벽시간연장,세포접충존활솔야명현강저,병출현쇠화현상.③면역조화감정결과 현시CD29、CD44표체기본양성,이CD34、CD45표체음성.결론 첩벽법비교용역분리배양MSCs,위조직공정적응용제공료족구적충자래원,MSCs유일정적증식능력단병불시무한적,이시수전대확증축점쇠화,인차료해화장악MSCs적생물학특성위진일보연구응용타하기출.
Objectives To investigate the feasibility of isolating and culturing bone marrow-derived mesenchymal stem cells (MSCs) in vitro by adherence method.Methods The experiment was conducted at institute of Urology ,First Affiliated Hospital of Nanchang University from January to June 2008.Neonate SD rats were selected. MSCs were isolated and cultured by the plastic adherence method in vitro and identified by CD29, CD34, CD44,CD45.Growth curve and the adhesive rate curve were drawn and biological properties of MSCs of different passages were analysed.Results ①The incubation period of primary generation was a bit longer. MSCs were round at the beginning and adhered to the wall in 24 hours. Most adherent cells were mainly monocytes and gradually changed into multiple cellular shapes 48 hours after primary culture. MSCs began to proliferate at day4 and coalesced 9 days later. ②The growth curve and the adhesive rate curve demonstrated that the growth velocity and adhesive rate were similar before passage 7.After passage 7, the growth velocity of MSCs was significantly dropped, the adherence rate also decreased, and apolexis emerged.③In vitro expanded MSCs are positive for CD29, CD44 and negative forCD34, CD45.Conclusions The adherence method can be manipulated conveniently. It is an ideal method to culture MSCs that provide sufficient source for tissue engineering. MSCs have limited proliferative capacity and showed apolexis with the increase of the passage. The study provides us with further understanding and mastering of biological characteristics and allows us to study the application of mesenchymal stem cells.
